Microscale Spatiotemporal Variation and Generation Mechanisms of Reactive Oxygen Species in the Rhizosphere of Ryegrass: Coupled Biotic-Abiotic Processes.

Reactive oxygen species (ROS) play key roles in soil biogeochemical processes, yet the occurrence and accumulation of ROS in the rhizosphere are poorly documented. Herein, we first developed a ROS-trapping membrane to in situ determine ROS in the ryegrass rhizosphere and then quantified the temporal and spatial variations of representative ROS (i.e., O2•─, H2O2, and •OH). Fluorescence imaging clearly visualized the production of ROS in the rhizosphere. Both O2•─ and H2O2 content increased first and then declined throughout the life cycle of ryegrass, while •OH concentration decreased continuously. Spatially, ROS contents remained at a relatively high level at 0-5 mm and then descended with increasing distance. The concentrations of ROS in different soils followed the order of black soil > latosol soil > yellow-brown soil > tier soil ∼ red soil. Analysis of soil properties suggested that both biotic factors (microbial community) and abiotic factors (Fe(II) and water-soluble phenols) played critical roles in ROS production. The combined processes, including Fe(II) and water-soluble phenol-mediated electron transfer, microbial community-driven extracellular O2•─ release, and Fe(II)/Fe(III) cycling, may be responsible for ROS production. These findings provide insights into ROS-associated rhizosphere effects and inspiration for the phytoremediation of pollutants and element cycling.

A random forest model for the classification of wheat and rye leaf rust symptoms based on pure spectra at leaf scale.

The pure spectra acquisition of plant disease symptoms is essential to improving the reliability of remote sensing methods in crop protection. The reflectance values read from the pure spectra can be used as valuable training data for development of algorithms designed for plant disease detection at leaf and canopy scale. The aim of this paper is to identify and distinguish spectrally the leaf rust symptoms caused by two closely related special forms (f. sp.) of Puccinia recondita f. sp. tritici on wheat and Puccinia recondita f. sp. recondita on rye at leaf scale. Spectral measurements were made with FieldSpec 3 spectrometer in the wavelength range of 350-2500 nm. The spectrometer was connected to a microscope by optical fiber. Raw spectra of uredinia, chlorotic discoloration, green leaves, senescent inoculated leaves and senescent uninoculated leaves of wheat and rye, all of which obtained for this study, were investigated with a view towards making an automized classification of plant species and their phases. The created Random Forest models were tested separately using pure spectra, and from these vegetation indices were derived as predictors. Three vegetation indices, namely CRI, PRI and GNDVI, appeared to be the most robust in terms of distinguishing uredinia from other symptoms on rye and wheat leaves. PRI, EVI, NDVI705, and GNDVI were the most suitable for distinguishing uredinia, chlorotic discoloration, and green leaf stages on rye. That tusk on wheat leaves can be recognized if seven indices (PRI, MSAWI, SAVI, NDVI, NDVI705, GNDVI and RVI) are used together. For the classification of all disease symptoms for both plant species, the most useful were wavelengths in the VIS range: 431-436, 696-703 and 646-686 nm. However, the ranges of SWIR wavelengths (1938, 1955) and NIR wavelengths (1099-1104) also have a high contribution to the discrimination accuracy of the model. In the classification of all disease symptoms, the most important vegetation indices were CRI, OSAVI, and GNDVI. Analysis of the results revealed the advantage of the model based on the selected spectral wavelengths (Hit Rate of 96.6%) in comparison with predictions based on vegetation indices alone (Hit Rate of 91.7%). Both approaches show the highly applicable character of utilizing high quality spectral products such as satellite images in reducing operational costs of crop protection.

Toxic metal phytoextraction potential and health-risk parameters of some cultivated plants when grown in metal-contaminated river sediment of Danube, near an industrial town.

Toxic metal phytoextraction potential of some higher plants, the white mustard (Sinapis alba L.), perennial rye grass (Lolium perenne L.) and also two cultivated plants, as green pea (Pisum sativum L. var. Rajnai törpe), radish (Raphanus sativus L. var. Szentesi óriás vaj), was studied in a field experiment, along the river Danube in close vicinity of an industrial town, Dunaújváros, Hungary. Soil/sediment and the various plant organs (leaves, stems and roots) were assessed for the contamination with some potentially toxic elements (PTE), such as the cadmium (Cd), nickel (Ni), copper (Cu), and zinc (Zn). It was found that Cd and Ni concentration was below, while the Cu and Zn elements were above the Hungarian permissible limits in each of the studied soil/sediment samples. Bioconcentration factor (BAF) was less than 1 in the shoot biomass of test plant samples and followed the order of Cu > Zn > Cd and Ni. Phytoremediation potential of selected test plants was found to be rather limited. The translocation factor (TF) was more than 1 for Cu and Zn elements, at each test plants. Cadmium was translocated into the leaves in case of the radish, only. Considering of the potential human daily intake of metals (DIM), it was less than 1 both for the adults and for the children. Health risk index (HRI) values of children, however, were higher than 1 for the Cd in case of radish, and for Zn and Cu in case of the pea. Results suggest that consumption of these plants grown in gardens of contaminated sediments can result in some risks for citizens in the industrial town of Dunaújváros. Further studies are required to identify appropriate plants with greater toxic metal phytoextraction potential.

Revealing the structural characteristics of lignin macromolecules from perennial ryegrass during different integrated treatments.

To reveal the structural characteristics and physicochemical properties of perennial ryegrass lignin, sequential alkali extractions or double ball-milling and enzymatic hydrolysis on the basis of ultrasonic and hydrothermal pretreatments were proposed in this study. Results revealed that sequential alkali extractions released 89.4% of original lignin from the ryegrass cell walls and 0.75-4.16% of associated carbohydrates as compared to the double ball-milling and enzymatic hydrolysis (96.0% and 18.39%). It was observed that the two types of lignin prepared were SGH-type and had different amounts of p-coumarates and ferulates, and primarily consisted of ß-O-4' linkages combined with minor amounts of ß-ß' and ß-5' linkages. Besides, alkali-soluble lignins exhibited relatively fewer ß-O-4' linkages, higher S/G ratios and H-type units, and abundant phenolic OH groups as compared to the double enzymatic lignin. Overall, the deeper investigation of the lignin structure of ryegrass will provide useful information for the efficient utilization of lignin macromolecules in biorefineries.

Dry matter productivity and bromatological quality of ryegrass genotypes cultivated in southern Brazil / [Produtividade de matéria seca e qualidade bromatológica de genótipos de azevém cultivados no Sul do Brasil]

The goal of this work was to assess the biomass production and bromatological quality of ryegrass genotypes in ten municipalities of the Western and North Plateau regions of the State of Santa Catarina, Brazil. The cultivars La Estanzuela 284 (diploid), Bar HQ, Barjumbo, INIA Escorpio, Potro, and Winter Star (tetraploids) were compared, distributed in a randomized block design, in which the municipalities constituted the blocks, with three replications. The cuts were performed when the plants reached 30cm, leaving a residue of 10cm. In three cuts, the cultivars Barjumbo and Bar HQ were the most productive, exceeding 4.6 t ha-1 of dry matter. In the places in which five cuts were performed, the production of these cultivars exceeded 7.3 t ha-1, placing them again ahead of the others. The average crude protein content in three cuts was greater than 25% in all cultivars. There was no difference between the genotypes in the content of neutral detergent fiber and total digestible nutrients. There was a significant correlation between quantitative and qualitative productive variables. The assessed cultivars represented good options for composing short-term or long-term winter-feeding systems, adjusted to the integration with annual crops or warm-season pastures.(AU)

O objetivo deste trabalho foi avaliar a produção de biomassa e qualidade bromatológica de genótipos de azevém, em dez municípios das regiões Oeste e Planalto Norte Catarinense, Estado de Santa Catarina, Brasil. Foram comparados os cultivares La Estanzuela 284 (diploide), Bar HQ, Barjumbo, INIA Escorpio, Potro e Winter Star (tetraploides), distribuídos em um delineamento blocos casualizados, em que os municípios constituíram os blocos, com três repetições. Os cortes foram realizados quando as plantas atingiram 30cm, deixando um resíduo de 10cm. Sob três cortes, os cultivares Barjumbo e Bar HQ foram os mais produtivos, ultrapassando 4,6 t ha-1 de matéria seca. Nos locais em que ocorreram cinco cortes, a produção destes cultivares superou 7,3 t ha-1, posicionando-os novamente à frente dos demais. O teor médio de proteína bruta em três cortes foi superior a 25% em todos os cultivares. Não houve diferença entre os genótipos no teor de fibra detergente neutro e de nutrientes digestíveis totais. Verificou-se correlação significativa entre variáveis produtivas quantitativas e qualitativas. Os cultivares testados representam boas opções para compor sistemas forrageiros hiberno-primaveris de curta ou longa duração, ajustando-se à integração com lavouras ou pastagens anuais de estação quente.(AU)

Integrated treatment of perennial ryegrass: Structural characterization of hemicelluloses and improvement of enzymatic hydrolysis of cellulose.

An integrated treatment coupling ultrasonic and hydrothermal pretreatments with sequential alkali post-extractions was performed to isolate and characterize hemicelluloses from perennial ryegrass and improve the enzymatic hydrolysis efficiency of cellulose. The yield, chemical composition, and structure of water-soluble and alkali-soluble hemicelluloses obtained from the hydrothermal supernatant and hydrothermally pretreated ryegrass as well as the enzymatic hydrolysis efficiency of cellulose were comprehensively investigated by gel permeation chromatograph, high-performance anion exchange chromatography, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectra. Results showed that more than 90 % of the original hemicelluloses in ryegrass were released during the integrated treatment and all hemicellulosic fractions obtained were mainly composed of ʟ-arabino-(4-O-methyl-ᴅ-glucurono)-ᴅ-xylans, galactoanrabinoxylans and ß-glucans. In addition, the effective removal of amorphous hemicelluloses and lignin significantly increased the cellulose enzymatic hydrolysis rate of ryegrass from 43.8 to 91.1 %. These results provided new insights into the collaborative utilization of hemicelluloses and cellulose in ryegrass.

Biostimulating effect of l-tryptophan on the yield and chemical and microbiological quality of perennial ryegrass (Lolium perenne) herbage and silage for ruminant.

BACKGROUND: This study aimed to evaluate the effect of l-tryptophan (l-TRP) used in the cultivation of Lolium perenne on the yield, and chemical and microbiological quality of its herbage and silage. l-Tryptophan was applied in doses of 5, 0.5, 0.05 kg ha-1 . The experiment was conducted with a control group (C) and a comparative control group (C+) with higher nitrogen fertilization. RESULTS: The dose of 5 kg ha-1 had a significant effect on herbage yield, which increased by 15% compared to group C and approximated the value achieved in group C+. The treatment with l-TRP caused a significant increase in water-soluble carbohydrate (WSC) content only in Tr5 (165 g kg-1 DM), which was reflected in a more beneficial course of fermentation, lower pH (4.59), and a higher sum of fermentation acids, including lactic acid (94.7 g kg-1 DM). Ryegrass treatment with a high l-TRP dose effectively reduced the loads of Clostridium spp. and fungi, and increased the count of Bacillus spp. The l-TRP significantly reduced N-NH3 content in Tr05 (98.6 g kg-1 TN) compared with C+ (123 g kg-1 TN) and butyric acid content in Tr05 (from 1.35 g kg-1 DM in the C to 0.38 g kg-1 DM). CONCLUSION: The most effective dose turned out to be a dose of 5 kg ha-1 , which allowed a higher yield and a better fermentation course to be achieved. This work presents the feasibility of using l-TRP to optimize nutrient consumption by Lolium perenne and ultimately to affect the quality of its silage as a feedstuff. © 2020 Society of Chemical Industry.

Comparison of cadmium uptake and transcriptional responses in roots reveal key transcripts from high and low-cadmium tolerance ryegrass cultivars.

Cadmium (Cd), which seriously affects plant growth and crop production, is harmful to humans. Previous studies revealed ryegrass (Lolium multiflorum Lam.) exhibits Cd tolerance, and may be useful as a potential hyperaccumulator because of its wide distribution. In this study, the physiological and transcriptional responses of two ryegrass cultivars [i.e., high (LmHC) and low (LmLC) Cd tolerance] to Cd stress were investigated and compared. The Cd tolerance of LmHC was greater than that of LmLC at various Cd concentrations. The uptake of Evans blue dye revealed that Cd-induced root cell mortality was higher in LmLC than in LmHC after a 12-h Cd treatment. Furthermore, the content and influx rate of Cd in LmLC roots were greater than in LmHC roots under Cd stress conditions. The RNA sequencing and quantitative real-time PCR data indicated that the Cd transport regulatory genes (ABCG37, ABCB4, NRAMP4, and HMA5) were differentially expressed between the LmLC and LmHC roots. This expression-level diversity may contribute to the differences in the Cd accumulation and translocation between LmLC and LmHC. These findings may help clarify the physiological and molecular mechanisms underlying ryegrass responses to Cd toxicity. Additionally, ryegrass may be able to hyperaccumulate toxic heavy metals during the phytoremediation of contaminated soil.

Determination of Arginine δ15N Values in Plant and Animal Proteins by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry.

Nitrogen (N) stable isotope techniques are widely used in ecology, archaeology, and forensic science to explore trophic relationships and provenances of organisms and materials, most widely using bulk δ15N values of whole organisms, tissues, or other materials. However, compound-specific isotope values can provide more diagnostic isotope "fingerprints" and specific information about metabolic processes. Existing techniques for nitrogen isotope analysis allow the determination of δ15N values of 14 amino acids (AAs), accounting for ca. 75% of plant protein and collagen N. The majority of remaining N is from arginine, comprising 16 and 14% of collagen and plant protein N, respectively. We therefore aimed to develop a method to detect arginine and determine its δ15N value (δ15NArg) by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS), to further contribute to the understanding of the metabolic routing of this important AA. We demonstrate that arginine, as its N-acetyl isopropyl ester, is amenable to GC analysis using a 15 m midpolarity DB-35 column, eluting with baseline resolution from other AAs. The recorded δ15N value by GC-C-IRMS was within the error of that of the underivatized compound determined by elemental analyzer-isotope ratio mass spectrometry (EA-IRMS). The newly developed GC-C-IRMS method was applied to modern plant protein and cattle collagen, enabling their δ15NArg values to be related to AA biosynthesis. Determination of archaeological cattle collagen δ15NArg values confirmed the suitability of this method to provide further insights into past diets and ecosystems. Bulk collagen δ15N value reconstruction including δ15NArg values better reflect the measured bulk values, as the isotopic ratio of 91% of collagen N can now be determined at the compound-specific level.

Fiber composition and digestibility of fodder in different ryegrass fields submitted to different harvesting management systems / [Composição da fibra e digestibilidade da forragem de diferentes cultivares de azevém submetidos a diferentes sistemas de manejo de colheita]

This essay aims to evaluate the composition of fiber in neutral detergent, digestibility of plant components, as well as ruminal disappearance rate of fodder dry matter from five ryegrass fields submitted to different cut systems for the production of haylage. The experimental design was the randomized blocks one, at a 5 x 3 factorial scheme, with five cultivars (cv.) of ryegrass Lolium multiflorum (cv. Barjumbo, cv. BarHQ, cv. Ribeye, cv. Potro and cv. Willians) associated with three harvesting management systems: 1- single harvesting system at a pre-floral stage; 2 - double harvesting system, with a cut at vegetative stage and another at pre-floral stage; and 3 - triple harvesting system, with a cut at vegetative stage, a cut at full vegetative stage and a cut at pre-floral stage, with three repetitions each. Similarities were noticed on the results of fodder digestibility of different fields, however, cv. Potro presented a higher ruminal disappearance rate after 48 hours, with 69.27%. Generally, the harvesting management system with the increase of ryegrass cut frequency determined increments on the percentage of lignin and the reduction in the concentration of total nutrients and dry matter digestibility of the resulting hylage.(AU)

O objetivo do presente trabalho foi avaliar a composição da fibra em detergente neutro, digestibilidade dos componentes da planta, bem como a taxa de desaparecimento ruminal da matéria seca da forragem de cinco cultivares de azevém submetidos a diferentes sistemas de cortes para produção de silagem pré-secada. O delineamento experimental foi em blocos ao acaso,, em um fatorial 5 x 3, sendo cinco cultivares de azevém Lolium multiflorum (cv. Barjumbo, cv. BarHQ, cv. Ribeye, cv. Potro e cv. Willians) associados a três sistemas de manejo de colheita: 1 - sistema com única colheita no estádio de pré-florescimento; 2 - sistema com uma colheita no estádio vegetativo e outra no estádio de pré-florescimento; e 3 - sistema com uma colheita no estádio vegetativo, uma no estádio pleno vegetativo e outra no estádio de pré-florescimento, com três repetições cada. Notou-se similaridade entre os resultados de digestibilidade da forragem entre os cultivares, porém o cv. Potro apresentou a maior taxa de desaparecimento ruminal após 48 horas, com 69,27%. De maneira geral, o sistema de manejo de colheita mostrou que o aumento do número de cortes do azevém determinou incrementos nos teores de lignina e redução na concentração de nutrientes digestíveis totais e da digestibilidade da matéria seca nas silagens pré-secadas resultantes.(AU)

Host Range Expansion of an Endemic Insect Herbivore is Associated With High Nitrogen and Low Fibre Content in Exotic Pasture Plants.

Endemic moth species of the genus Wiseana spp. (Hepialidae) have become serious pests of introduced pasture plants in New Zealand. The original native host plants of these moths have not been confirmed. This study investigated the performance (survival, development time, weight gain) of three Wiseana species on seven putative host plants: five native and two exotic species. The aim was to identify native hosts for the three Wiseana species and to compare their performance on native plants and exotic pasture plants. The chemical composition of the seven putative host plants was investigated to compare native and exotic plant chemistries, and to test for associations between plant characteristics and performance of selected Wiseana species. Carbon, nitrogen, silica and fibre contents were measured for each plant species; primary metabolite composition was determined by gas chromatography-mass spectrometry. For the three moth species, increased survival and weight gain were significantly associated with high nitrogen and low fibre contents in one exotic host plant, white clover (Trifolium repens), although one species, W. umbraculata, did not complete development to adult on any of the plants tested, including clover. Two exotic plants (T. repens, Lolium perenne × Lolium multiflorum), and two native plants (Aciphylla squarrosa and Festuca actae) supported W. copularis development to the adult stage, but only one exotic (T. repens) and one native (F. actae) species supported complete development of W. cervinata. Exotic and native plant species had distinct metabolite profiles, but there was no significant association between metabolite composition and Wiseana performance. We conclude that W. copularis and W. cervinata, but not W. umbraculata, have expanded their host range, because of their ability to use both native and new hosts. No evidence was found for a host shift, i.e., a loss of performance on the ancestral host compared with the new host.

Metal immobilization and nitrate reduction in a contaminated soil amended with zero-valent iron (Fe0).

Technologies based on zero-valent iron (Fe0) are increasingly being used to immobilize metals in soils and remove metals and nitrate from waters. However, the impact of nitrate reduction on metal immobilization in metal contaminated soils has been poorly investigated so far. Here, different concentrations of Fe0 filings (1%, 2% and 5%; wt%) were applied to a metal contaminated soil. The resulting nitrate reduction and metal (Cd and Zn) immobilization was investigated using a column leaching experiment for 12 weeks. Corrosion of Fe0 filings and precipitation of Fe oxyhydroxydes (FeOOH) on the surfaces of the filings were observed using SEM-EDS and EMPA-WDS at the end of the experiment. Compared to the untreated soil, total nitrate amounts released were lowered by 47%, 59% and 87% in the presence of 1%, 2% and 5% of Fe0, respectively. Concomitantly with nitrate reduction, Cd and Zn concentrations in leachates were strongly alleviated in the presence of Fe0, which was partly attributed to the rise of soil pH subsequent to nitrate reduction. More importantly, biotests with Lupinus albus L. revealed that the mechanisms involved in metal immobilization are stable to root-induced acidification. However, Fe0 was not efficient to reduce Cd concentration in Lolium multiflorum Lam., indicating that root processes other than acidification may re-mobilize metals.

Mycelial biomass and concentration of loline alkaloids driven by complex population structure in Epichloë uncinata and meadow fescue (Schedonorus pratensis).

Many efforts have been made to select and isolate naturally occurring animal-friendly Epichloë strains for later reinfection into elite cultivars. Often this process involves large-scale screening of Epichloë-infected wild grass populations where strains are characterized and alkaloids measured. Here, we describe for the first time the use of genotyping-by-sequencing (GBS) on a collection of 217 Epichloë-infected grasses (7 S. arundinaceum, 4 L. perenne, and 206 S. pratensis). This genotyping strategy is cheaper than complete genome sequencing, is suitable for a large number of individuals, and, when applied to endophyte-infected grasses, conveniently genotypes both organisms. In total, 6273 single nucleotide polymorphisms (SNPs) in the endophyte data set and 38 323 SNPs in the host data set were obtained. Our findings reveal a composite structure with three distinct endophyte clusters unrelated to the three main S. pratensis gene pools that have most likely spread from different glacial refugia in Eurasia. All three gene pools can establish symbiosis with E. uncinata. A comparison of the endophyte clusters with microsatellite-based fingerprinting of the same samples allows a quick test to discriminate between these clusters using two simple sequence repeats (SSRs). Concentrations of loline alkaloids and mycelial biomass are correlated and differ significantly among the plant and endophyte subpopulations; one endophyte strain has higher levels of lolines than others, and one specific host genotype is particularly suitable to host E. uncinata. These findings pave the way for targeted artificial inoculations of specific host-endophyte combinations to boost loline production in the symbiota and for genome association studies with the aim of isolating genes involved in the compatibility between meadow fescue and E. uncinata.

Research Note: Effect of diet with different proportions of ryegrass on breast meat quality of broiler geese.

This study was conducted to determine the effects of diet with different proportions of ryegrass on breast meat quality of geese. In total, 240 healthy male Yangzhou geese (28-day-old) with similar body weight were divided randomly into 4 diet groups (control group: fed commercial diets; treatment groups I, II, and III: fed ryegrass and commercial diet in the ratios of 1.5:1, 2:1, and 3:1, respectively), the birds being fed from the age of 29 to 70 D. The results shows that the body weights of 70-day-old geese of treatment groups II and III were lower than those in the control group, whereas those of geese of treatment group I were similar to those of the control group. The contents of flavor amino acid and total (essential) amino acids in treatment groups I and II were higher than those in treatment group III (P < 0.05). In addition, grass supplementation reduced saturated fatty acid content and increased that of omega-3 (n-3) polyunsaturated fatty acids, relative to the control group (P < 0.05). Finally, among the 6 minerals analyzed in breast muscle, differences existed in Zn, Se, and Cu contents among the geese fed with different proportions of ryegrass. Zn content of geese from treatment groups II and III was significantly higher than that of those of the control group; Cu content was lower with grass intake and was significantly higher in the control group than in treatment group III; Se content was significantly higher in the control group than in both groups II and III (all at P < 0.05). The results from this study indicated that geese fed with low proportions of ryegrass (1.5:1 or 2:1) showed good growth performance and increased total (essential) amino acid, flavor amino acid, n-3 polyunsaturated fatty acid, and Zn content in meat, which had a certain guiding value for the production of high-quality goose meat under intensive feeding conditions.

Heavy Metal Uptake of Lettuce and Ryegrass from Urban Waste Composts.

Interest in using urban waste composts as amendments in urban agriculture is growing nowadays. However, concerns about the potential transference of pollutants present in urban waste to the food chain are very relevant when they are recycled for food or animal feed production. Thus, for the safe use of urban waste composts, it has to be assured that no risk of metal transference to plants from compost exists. In this work, the transference of heavy metals from urban waste composts to plants has been studied in an experiment with lettuce and Italian ryegrass, grown in substrates based on five metal-rich composts and a manure vermicompost (included for comparison). A two-month pot experiment was performed under controlled light and temperature conditions, and plant growth and uptake of Cu, Pb, Cd and Zn were analyzed. For both species, the concentration of metals in plant tissue followed the sequence Zn > Cu >> Pb ≈ Cd, the same as the metal concentrations in four out of the five composts. Plant concentrations of Zn, Cu and Cd increased with their concentrations in compost, whereas this relation was not observed for Pb. The ratio between element concentration in plant and compost were much higher for Zn, Cd and Cu than for Pb, showing the lower bioavailability of Pb with respect to other metals.

Contrasting Effects of Grass - Endophyte Chemotypes on a Tri-Trophic Cascade.

Systemic grass-endophytes of the genus Epichloë symbiotically infect the above-ground plant parts of many grass species, where they produce alkaloids in a grass- and endophyte-specific manner that are toxic or deterrent to herbivores. An increasing number of studies show cascading negative effects of endophyte-derived alkaloids that extend to higher trophic levels, harming beneficial insects, including those that control aphid populations. Lacewings are one of the major biological aphid controls, and are especially resistant to insecticides and pollutants, but their susceptibility to endophyte infection in the food chain has never been studied. Our study found variability in aphid population growth depending on the endophyte-grass chemotype, where aphid population growth was lowest on chemotypes known for producing high amounts of loline alkaloids. We also showed that larval and pupal development and mortality of the Common Green Lacewing (Chrysoperla carnea) was, in a non-choice experiment, not affected by endophyte infection in the food chain. This is a first indication that lacewings might be resistant to endophyte-derived alkaloids and could be robust biocontrol agents when applied together with endophyte-infected grass, possibly replacing chemical pesticides.

Application of co-composted biochar significantly improved plant-growth relevant physical/chemical properties of a metal contaminated soil.

A woody-biochar was added to waste biomass during a composting process. The resulting compost-char was amended to a metal contaminated soil and two plant species, L. perenne and E. sativa, were grown in a pot experiment to determine 1) plant survival and stress factors, 2) uptake of metals to plants and, 3) chemical characteristics of sampled soils and pore waters. Compost supplemented with biochar after the composting process were also tested, as well as a commercially available compost, for comparison. Co-composting with biochar hastened the composting process, resulting in a composite material of reduced odour, increased maturity, circum-neutral pH and increased moisture retention than compost (increase by 3% of easily removable water content). When amended to the soil, CaCl2 extractable and pore water metals s were reduced by all compost treatments with little influence of biochar addition at any tested dose. Plant growth success was promoted furthest by the addition of co-composted biochar to the test soil, especially in the case of E. sativa. For both tested plant species significant reductions in plant metal concentrations (e.g. 8-times for Zn) were achieved, against the control soil, by compost, regardless of biochar addition. The results of this study demonstrate that the addition of biochar into the composting process can hasten the stability of the resulting compost-char, with more favourable characteristics as a soil amendment/improver than compost alone. This appears achievable whilst also maintaining the provision of available nutrients to soils and the reduction of metal mobility, and improved conditions for plant establishment.

Milk and meat fatty acids from sheep fed a plantain-chicory mixture or a grass-based permanent sward.

Plantain and chicory are interesting forage species since they present good nutritional quality and are more resistant to drought than many temperate grasses. The fatty acid (FA) profile in milk and meat is related to a growing concern for the consumption of healthy foods, that is, with a lower content of saturated FA, higher polyunsaturated FA (PUFA) and a favourable n-6 : n-3 FAs ratio. Our objective was to evaluate the FA content in ewe's milk and lamb's meat fed a plantain-chicory mixture (PCH) or a grass-based permanent sward (GBS) dominated by perennial ryegrass. Eighteen Austral ewes in mid-lactation were allocated to PCH and GBS treatments. Milk samples were obtained during September (spring). Thirty weaned lambs were finished on both treatments from November to December (7 weeks), slaughtered and their meat sampled. Fat from milk and meat samples was extracted and stored until analysed by gas chromatography. Milk fat from GBS was higher than from PCH (P < 0.05) in C18:0 (11 385 v. 5874 mg/100 g FA), 9c-18:1 (15 750 v. 8565 mg/100 g FA), 11 t-18:1 (4576 v. 2703 mg/100 g FA) and 9c,11 t-18:2 (1405 v. 921 mg/100 g FA) and lower in 18:2n-6 (827 v. 1529 mg/100 g FA) and 18:3n-3 (943 v. 1318 mg/100 g FA) FA. Total mono-unsaturated FA was higher in GBS than PCH (P < 0.05). Meat fat from PCH swards presented a higher (P < 0.05) content than GBS for 18:2n-6 (46.8 v. 28.2 mg/100 g FA), linolenic (24.6 v. 14.2 mg/100 g FA), polyunsaturated FA (119.7 v. 73.4 mg/100 g FA), n-6 (65.9 v. 40.8 mg/100 g FA) and n-3 (53.8 v. 32.5 mg/100 g FA), respectively. No effect of treatment (P > 0.05) was detected for 9c-18:1 (283.9 v. 205.8 mg/100 g FA), 11 t-18:1 (26.2 v. 19.3 mg/100 g FA) and 9c,11 t-18:2 (10.1 v. 7.6 mg/100 g FA), for PCH and GBS. These results suggest that grazing a PCH mixture results in a higher concentration of PUFA in ewes' milk and in lambs' fat, as compared to a GBS sward.

Effect of varying fermentation conditions with ensiling period and inoculum on photosynthetic pigments and phytol content in Italian ryegrass (Lolium multiflorum Lam.) silage.

This study aimed to investigate the effect of an ensiling period (Experiment 1) and adding lactic acid bacteria (LAB, Experiment 2) on the changes in carotenoid, chlorophyll, and phytol in ensiled Italian ryegrass (IR, Lolium multiflorum Lam.). In Experiment 1, the IR herbage ensiled into plastic bags was analyzed for the contents of photosynthetic pigments and phytol over a 5-week period. During the ensiling process, the ß-carotene content decreased (p < .05), whereas the lutein content did not change. Although the chlorophyll content decreased (p < .05) after ensiling, the phytol content barely changed until week 5. In Experiment 2, IR herbage was ensiled without additive, as a Control, or with LAB for 60 days. The pH was lower (p < .05) and lactic acid content was higher (p < .05) for the LAB silage than for the Control. The chlorophyll content in silage was not affected by the LAB; however, the ß-carotene content was higher (p < .05) for the LAB silage than for the Control. Phytol and lutein contents in the herbage did not change after ensiling. These results indicate that phytol and lutein in IR herbage can be preserved well in silage, irrespective of their fermentation condition.

Perennial Ryegrass Alkaloids Increase Respiration Rate and Decrease Plasma Prolactin in Merino Sheep under Both Thermoneutral and Mild Heat Conditions.

A study was undertaken to determine the effects of feeding two levels of perennial ryegrass alkaloids (nil vs. moderate) under two climatic conditions. Alkaloids were fed via endophyte-infected perennial ryegrass seed and hay. Twenty-four Merino ewe weaners (six months, initial BW 32 ± 1.7 kg) were used in a study that lasted for 21 days after 14 days of adaptation. Sheep were fed either a control or alkaloid (Alk, 110 µg/kg LW ergovaline and 75 µg/kg LW lolitrem B) supplemented diet. Sheep were exposed to either constant thermoneutral (TN, 21-22 °C, 49% RH) or mildly heated (HS, 33 °C 1000-1500 h, 28% relative humidity) conditions. Dietary Alk and HS reduced dry matter intake (DMI) (p < 0.001, p = 0.02, respectively) with the combination of both reducing DMI by 42%. Reductions in DMI resulted in a lower daily gain in the Alk treatment (p < 0.001). Feed digestibility was reduced in the combined treatment (p = 0.03). Rectal temperature, respiration rate, and skin temperature increased in the Alk treatment. Plasma prolactin concentrations were decreased by Alk and increased by mild HS. The data indicate that production is compromised in the presence of Alk and mild HS, with this effect being exacerbated by a combination of both.